比较RP-HPLC法与荧光偏振免疫法测定苯巴比妥、卡马西平、苯妥英钠在癫痫患者的血药浓度

目的 建立同时测定人血清苯巴比妥、卡马西平、苯妥英钠(抗癫痫药)浓度的RP-HPLC法,并与荧光偏振免疫法测定方法比较.方法 以乙酸乙酯-二氯甲烷(4:1)为提取溶剂、氯硝西泮为内标,固定相为Diamonsil C18柱(250mm×4.6 mm,5 μm),流动相为乙腈-水(35:65),紫外检测波长285 nm,流量1.0 mL·min-1;收集患者服药后稳态谷浓度样本,分别用2种方法测定,考察2种方法的相关性.结果 3种药物均具有良好的线性范围,日内、日间RSD均小于8%;苯妥英钠、苯巴比妥2种方法的测定值无统计学差异(P>0.05);而卡马西平2种测定方法比较有统计学差异(P<0.01),卡马西平RP-HPLC法测定值较低.结论 RP-HPLC法适用于治疗药物浓度监测.     

RP-HPLC法同时测定人血浆中多种抗癫痫药物浓度

目的建立RP-HPLC法同时测定人血浆中苯巴比妥、苯妥英钠、卡马西平浓度,为临床癫痫患者个体化治疗方案的实施提供指导。方法血清样品经乙醚提取后Agilent ZORBAX SB-C 18色谱柱(150mm×4.6mm,5μm)进行分析,流动相:甲醇-水(40∶60);流速:1.0mL·min^-1;检测波长:220nm;柱温:30℃;三种药物互为内标。结果在一定范围内(苯巴比妥2~98mg·L^-1,r=0.9990;苯妥英钠1.005~50.25mg·L^-1,r=0.9992,卡马西平0.4~30mg·L^-1,r=0.9995)各药物与内标峰面积比与浓度呈良好的线性关系。相对回收率分别为101.10%、102.21%和100.12%;低、中、高浓度质控品回收率均>90%(n=5),日内、日间变异均<10%(n=5)。结论本方法操作简单、成本低、精密度及回收率均满足生物样本分析要求,结果稳定可靠,适合在基层医院开展。     

UPLC-MS/MS同时测定人血浆中7种抗癫痫药物及1种活性代谢产物的血药浓度

目的 建立超高效液相色谱-质谱法(UPLC-MS/MS)同时测定人血浆中丙戊酸钠、卡马西平、地西泮、苯妥英钠、苯巴比妥、拉莫三嗪和奥卡西平等7种抗癫痫药物,以及1种活性代谢产物(10,11-二氢-10-羟基卡马西平)的方法,并应用于临床检测.方法 采用C18色谱柱(2.1 mm×l00 mm·1.7 μm),2 mmo...     

HPLC法同时测定5种抗癫痫药的血清药物浓度

目的建立同时测定拉莫三嗪、苯巴比妥、苯妥英、卡马西平和氯硝西泮血清浓度的HPLC法。方法血清样品经甲醇沉淀蛋白后直接进样分析,色谱柱为Waters C18(5μm,4.6×250mm)柱,流动相为甲醇-水(55∶45),检测波长235nm。结果在一定浓度范围内(拉莫三嗪:1.3~50.0μg/m L;苯巴比妥:2.5~100.0μg/m L;苯妥英:2.2~70.0μg/m L;卡马西平:1.8~35.0μg/m L;氯硝西泮:2.5~80.0μg/m L),各药物的峰面积与浓度呈良好的线性关系。低、中、高浓度的质控样品回收率均高于95%,相对标准差(RSD)均小于10%。结论本方法操作简便,结果稳定可靠,适用于临床进行血药浓度监测。     

HPLC法测定血清中苯巴比妥和卡马西平的浓度

目的建立人血清中苯巴比妥、卡马西平浓度的高效液相色谱含量测定方法。方法采用ODS-Hypersil(4.6 mm×100 mm,5μm)色谱柱,以甲醇∶水(55∶45)为流动相,紫外检测波长为254nm,以苯妥英为内标。结果苯巴比妥在2.5~40μg.mL-1浓度范围内线性关系良好(r=0.9996),平均回收率为100.69%,RSD为1.8%;卡马西平在1.25~20μg.mL-1浓度范围内线性关系良好(r=0.9995),平均回收率为98.67%,RSD为2.1%。日内和日间RSD均不大于3%。结论本方法准确、快速、简便,可作为苯巴比妥和卡马西平血药浓度监测的常规方法。     

HPLC法同时测定血清中苯巴比妥、苯妥英和卡马西平的浓度

目的 :建立快速测定血清中苯巴比妥、苯妥英和卡马西平浓度的方法。方法 :色谱柱DiamonsilC18(4 .6mm× 2 5 0mm ,5 μm) ,流动相甲醇∶水∶乙腈 (4 0∶4 5∶15 ) ,流速1.2ml·min-1,检测波长 2 4 0nm ,内标硝西泮 ,柱温 4 0℃。结果 :提取回收率为 84 .4～ 89.2 % ,日内和日间RSD均 <7.9% ;标准曲线相关性良好。对 10 4例患者作血清浓度监测 ,临床效果满意。结论 :本方法可作为苯巴比妥、苯妥英和卡马西平的血药浓度监测的常规方法。     

高效液相色谱法同时测定苯巴比妥、苯妥英钠和卡马西平血药浓度

目的：建立同时测定血清中苯巴比妥,苯妥英钠和卡马西平的浓度的高效液相色谱（ＨＰＬＣ）,方法：血清乙酸乙酯提取浓集后进样,采用Ｃ１８柱,以甲醇－水（５０：５０）为流动相,苯乙酮为内标物,在波长２１０ｎｍ处测定。     

638例次抗癫痫药血清浓度监测结果及其与临床疗效关联性分析

目的:为临床抗癫痫药物(AEDs)的合理使用提供科学参考。方法:对采用荧光偏振免疫(FPIA)法对丙戊酸钠、卡马西平、苯妥英钠、苯巴比妥进行血清浓度监测结果进行回顾性统计分析。结果:共监测638例次,359例次(56.27%)血药浓度位于治疗窗内,202例次(31.66%)低于治疗窗,70例次(10.97%)高于治疗窗,7例次(1.10%)未检出血清浓度;各AEDs血清浓度位于治疗窗的比例分别为卡马西平78.65%、苯巴比妥60.00%、丙戊酸钠57.48%、苯妥英钠23.08%,其中卡马西平、丙戊酸钠、苯妥英钠血清浓度位于治疗窗内癫痫治疗有效率分别为72.86%、90.57%、94.44%,明显高于其他浓度范围(χ2卡马西平=6.324,P卡马西平=0.012;χ2丙戊酸钠=122.782,P丙戊酸钠=0.000;χ2苯妥英钠=19.584,P苯妥英钠=0.000),而苯巴比妥在各浓度范围有效率差异无统计学意义(χ2=3.403,P=0.065);53例次联合用药中,12例次血清浓度在治疗窗内,占联合用药例次的22.64%。结论:对癫痫患者进行血清浓度监测,实施个体化给药,对促进抗癫痫药物安全、有效、合理使用具有重要意义。     

高效液相色谱法测定人血清中卡马西平及环氧化卡马西平浓度

目的建立测定人血清中卡马西平及其代谢物10,11-环氧化卡马西平浓度的方法.方法采用高效液相色谱法,血样用乙醚提取,以艾司唑仑为内标,色谱柱为Kromasil C18柱(150 mm ×4.6 mm,5μm),流动相为乙腈-水(1:2),检测波长215 nm,流速1.2ml·min-1.结果血清中卡马西平、环氧化卡马西平线性范围分别为1.0～20.0 mg·L-1(r=0.999 9)、0.1～5.0 mg·L-1(r=0.999 4),平均回收率接近100%,日内、日间RSD均小于5%.结论本法快速、简便、准确,适用于临床常规监测需要.     

苯巴比妥和苯妥英钠及卡马西平血药浓度测定

目的：建立一种简便可行的反相高效液相色谱法(RP HPLC)，一步同时测定苯巴比妥、苯妥英钠、卡马西平的血药浓度。方法：患者血清经二氯甲烷提取后在C18柱上分析，流动相为甲醇∶水(57∶43)，柱温30℃，紫外检测波长254 nm，流速0.8 mL·min 1。结果：线性范围分别为2.5～40，2.5～40，1.25～20 μg·mL 1，日内和日间RSD均小于10.0%(n＝5)。结论：该法简便、稳定，用于苯巴比妥、苯妥英钠、卡马西平的血药浓度监测，效果良好。     

Efficacy of gut lavage,hemodialysis, and hemoperfusion in the therapy of paraquat or diquat intoxication

Clinical and in vitro investigations were carried out to test the efficacy of gut lavage, hemodialysis, and hemoperfusion in the treatment of poisoning with paraquat or diquat. In a patient suffering from diquat intoxication 130 times more diquat was removed by gut lavage 30 h after ingestion than was removed by complete aspiration of the gastric contents.Determination of in vitro clearances for paraquat and diquat by hemodialysis showed that, at serum concentrations of 1–2 ppm, such as are frequently encountered in poisoning in man, toxicologically relevant quantities of herbicide cannot be removed from the body. At a concentration of 20 ppm, on the other hand, hemodialysis proved to be effective, the clearance being 70 ml/min at a blood flow rate of 100 ml/min. The efficacy of hemoperfusion with coated activated charcoal was on the whole better. Especially at concentrations around 1–2 ppm, the clearance values for hemoperfusion were some 5–7 times higher than those for hemodialysis.In a patient suffering from paraquat poisoning, both hemodialysis as well as hemoperfusion were carried out. The in vitro results could be confirmed: At serum concentrations of paraquat less than 1 ppm no clearance could be obtained by hemodialysis while by hemoperfusion with activated charcoal quite high clearance values were measured and the serum level dropped down to zero.

---

Zusammenfassung Klinische Untersuchungen und Laboratoriumsversuche wurden durchgeführt, um die Wirksamkeit von Darmspülung, Hämodialyse und Hämoperfusion bei Paraquat- und Deiquat-Vergiftungen zu prüfen.Bei einem Patienten wurde 30 Std nach Deiquat-Aufnahme durch Darmspülung 130mal mehr Deiquat entfernt als durch vollständige Aspiration des Mageninhaltes. In vitro-Versuche ergaben, daß bei Blutserumkonzentrationen von 1–2 ppm, die bei Vergiftungen oft gemessen werden, durch Hämodialyse keine toxikologisch relevanten Paraquat- oder Deiquat-Mengen entfernt werden können. Dagegen erwies sich die Hämodialyse bei 20 ppm und einer Blutumlaufgeschwindigkeit von 100 ml/min mit einer Clearance von 70 ml/min als wirksam. Die Hämoperfusion mit beschicheter Aktivkohle war in diesen Versuchen aber eindeutig überlegen, denn insbesondere bei Konzentrationen um 1–2 ppm waren die Clearance-Werte 5–7mal höher als bei der Hämodialyse.Die in vitro-Ergebnisse wurden bei einem Patienten mit einer Paraquat-Vergiftung bestätigt: Bei Konzentrationen unter 1 ppm war die Hämodialyse wirkungslos, während durch Hämoperfusion relativ hohe Clearance-Werte erreicht wurden, so daß der Serumspiegel rasch unter die Nachweisgrenze abfiel.

     

In vitro studies on sterically stabilized liposomes (SL) as enzyme carriers in organophosphorus (OP) antagonism

This study describes a new approach for organophosphorous (OP) antidotal treatment by encapsulating an OP hydrolyzing enzyme, OPA anhydrolase (OPAA), within sterically stabilized liposomes. The recombinant OPAA enzyme was derived from Alteromonas strain JD6. It has broad substrate specificity to a wide range of OP compounds: DFP and the nerve agents, soman and sarin. Liposomes encapsulating OPAA (SL)* were made by mechanical dispersion method. Hydrolysis of DFP by (SL)* was measured by following an increase of fluoride ion concentration using a fluoride ion selective electrode. OPAA entrapped in the carrier liposomes rapidly hydrolyze DFP, with the rate of DFP hydrolysis directly proportional to the amount of (SL)* added to the solution. Liposomal carriers containing no enzyme did not hydrolyze DFP. The reaction was linear and the rate of hydrolysis was first order in the substrate. This enzyme carrier system serves as a biodegradable protective environment for the recombinant OP-metabolizing enzyme, OPAA, resulting in prolongation of enzymatic concentration in the body. These studies suggest that the protection of OP intoxication can be strikingly enhanced by adding OPAA encapsulated within (SL)* to pralidoxime and atropine.     

Aquatic Insects and Trace Metals: Bioavailability,Bioaccumulation, and Toxicity

Abstract

The uptake of metals from food and water sources by insects is thought to be additive. For a given metal, the proportions taken up from water and food will depend both on the bioavailable concentration of the metal associated with each source and the mechanism and rate by which the metal enters the insect. Attempts to correlate insect trace metal concentrations with the trophic level of insects should be made with a knowledge of the feeding relationships of the individual taxa concerned. Pathways for the uptake of essential metals, such as copper and zinc, exist at the cellular level, and other nonessential metals, such as cadmium, also appear to enter via these routes. Within cells, trace metals can be bound to proteins or stored in granules. The internal distribution of metals among body tissues is very heterogeneous, and distribution patterns tend to be both metal and taxon specific. Trace metals associated with insects can be both bound on the surface of their chitinous exoskeleton and incorporated into body tissues. The quantities of trace meals accumulated by an individual reflect the net balance between the rate of metal influx from both dissolved and particulate sources and the rate of metal efflux from the organism. The toxicity of metals has been demonstrated at all levels of biological organization: cell, tissue, individual, population, and community. Much of the literature pertaining to the toxic effects of metals on aquatic insects is based on laboratory observations and, as such, it is difficult to extrapolate the data to insects in nature. The few experimental studies in nature suggest that trace metal contaminants can affect both the distribution and the abundance of aquatic insects. Insects have a largely unexploited potential as biomonitors of metal contamination in nature. A better understanding of the physico-chemical and biological mechanisms mediating trace metal bioavailability and exchange will facilitate the development of general predictive models relating trace metal concentrations in insects to those in their environment. Such models will facilitate the use of insects as contaminant biomonitors.     

Steroidal sapogenin contents in some domestic plants

In order to find out the values of the steroid resources for the future use. the compositions and contents of steroidal sapogenins from 13 domestic plants have been investigated. As a result,Dioscorea nipponica, D. quinqueloba andSmilax china were found to have large amount of diosgenin. And pennogenin inTrillium kamtschaticum andParis verticillata, yuccagenin inAllium fistulosum, hecogenin inAgave americana and neochlorogenin inSolanum nigum were appeared to be major steroidal sapogenins.     

Alzheimer’s disease – current trends in basic and clinical research. Highlights from the 16th ECNP

Advances in the molecular biological knowledge of neuronal nicotinic acetylcholine receptors (nAChRs) have led to a growing interest by the pharmaceutical industry in the development of novel compounds that selectively modulate nAChR function. The ability of (-)-nicotine, an activator of nAChRs, to enhance attentional aspects of cognition in animals and humans, to exert neuroprotective and anxiolytic-like effects, and presumably to mediate the negative correlation between smoking and Alzheimer's (and Parkinson's) Disease, has focused interest on the potential therapeutic utility of modulators of nAChR function for treatment of some of the deficits associated with these progressive, neurodegenerative conditions. Numerous compounds are known which activate nAChRs and which might serve as lead compounds toward the development of such agents. The pharmacologic diversity of neuronal nAChR subtypes suggests the possibility of developing selective compounds which would have more favourable side-effect profiles than existing agents. This broader class of agents, collectively called cholinergic channel modulators (ChCMs), is anticipated to encompass compounds which would have more favourable side-effect profiles than existing agents, which generally exhibit low selectivity. This selectivity may be achieved by preferentially activating some subtypes of nAChRs (i.e., Cholinergic Channel Activators, ChCAs) or inhibiting the function of other subtypes (Cholinergic Channel Inhibitors, ChCIs). An overview of the biology of nAChRs and the rationale for the use of ChCMs for the treatment of dementia related to neurodegenerative diseases are presented, followed by a discussion of lead compounds and compounds under consideration for clinical evaluation.